Brevilin A Inhibits VEGF-Induced Angiogenesis through ROS-Dependent Mitochondrial Dysfunction

Figure 3

The effects of Brevilin A on apoptosis and PI3K/AKT signaling pathway in human endothelial cells. After the BA application for 12 h, apoptotic phenotypes and signaling pathways were determined in HUVECs cultured in the presence of VEGF (10 ng/mL). (a) Morphological changes of confluent HUVEC cultures observed under a phase-contrast microscope (scale bars: 100 μm; ). (b) Flow cytometry evaluated different apoptotic phases via using AV-FITC/PI staining reagents (). (c and d) Bar graphs quantifying the relative distribution of HUVECs in early stage (AV⁺) or late apoptotic stages (AV⁺/PI⁺) (). Western blotting was used to determine (e, f) the protein expression of Bax, Bcl-2, and cleaved caspase-3 (). (g) HUVECs lysates were harvested and further subjected to caspase-3 activity determination (). Western blotting was used to determine (h, i) the protein expression of phosphorylation levels of PI3K and AKT (). The graphs summarize the data from at least three times independent experiments. Data are shown as the . One-way ANOVA with Bonferroni post hoc test and , , and versus solvent (Sol).

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